Human Phospho-GSK-3 beta (S9) Antibody

  
  • Species Reactivity
    Human
  • Specificity
    Detects human GSK-3 beta when phosphorylated at S9.
  • Source
    Monoclonal Mouse IgG1 Clone # 609739
  • Purification
    Protein A or G purified from hybridoma culture supernatant
  • Immunogen
    Phosphopeptide containing the human GSK-3 beta S9 site
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    1 µg/mL
    See below
  • CyTOF-ready
    Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.
  • Immunocytochemistry
    8-25 µg/mL
    See below
  • Intracellular Staining by Flow Cytometry
    2.5 µg/106 cells
    See below
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Data Examples
Detection of Human Phospho-GSK‑3 beta (S9) by Western Blot. Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line untreated (-) or treated (+) with 200 nM PMA for 20 minutes. PVDF Membrane was probed with 1 µg/mL of Human Phospho-GSK‑3 beta (S9) Monoclonal Antibody (Catalog # MAB25062) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for Phospho-GSK‑3 beta (S9) at approximately 46 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Immunocytochemistry
Phospho-GSK‑3 beta (S9) in HeLa Human Cell Line. GSK‑3 beta phosphorylated at S9 was detected in immersion fixed HeLa human cervical epithelial carcinoma cells, unstimulated (lower panel) or stimulated (upper panel) with PMA, using Mouse Anti-Human Phospho-GSK‑3 beta (S9) Monoclonal Antibody (Catalog # MAB25062) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Intracellular Staining by Flow Cytometry
Detection of Phospho-GSK-3 beta (S9) in HeLa Human Cell Line by Flow Cytometry. HeLa human cervical epithelial carcinoma cell line untreated (open histogram) or treated with 120 ng/mL PMA for 15 minutes (filled histogram) was stained with Human Phospho-GSK‑3 beta (S9) Monoclonal Antibody (Catalog # MAB25062) or isotype control antibody (Catalog # MAB002, not shown), followed by Fluorescein-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0103B). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with saponin.
Preparation and Storage
  • Reconstitution
    Sterile PBS to a final concentration of 0.5 mg/mL.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • Stability & Storage
    Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
    • 12 months from date of receipt, -20 to -70 °C as supplied.
    • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
    • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: GSK-3 beta

Glycogen Synthase Kinase-3 (GSK-3) is a serine/threonine kinase initially identified asan inhibitor of glycogen synthase. Two isoforms (GSK-3 alpha and GSK-3 beta ) share 85% amino acid identity. GSK-3 beta, inhibited by phosphorylation at S9 by Akt, is involved in energy metabolism, body pattern formation, and neuronal cell development.

  • Long Name:
    Glycogen Synthase Kinase 3 beta
  • Entrez Gene IDs:
    2932 (Human); 56637 (Mouse); 84027 (Rat)
  • Alternate Names:
    EC 2.7.11; EC 2.7.11.26; glycogen synthase kinase 3 beta; glycogen synthase kinase-3 beta; GSK3 beta; GSK-3 beta; GSK3B; GSK3beta isoform
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